8-羟基大豆苷元的生物活性研究进展

8-羟基大豆苷元(8-hydroxydaidzein)是在一些发酵豆制品中,由大豆苷元(daidzein)在曲霉菌羟基化酶作用下发生C8羟基化反应的产物。该文对8-羟基大豆苷元的DPPH自由基清除活性、抗氧化活性、酪氨酸酶抑制活性以及醛糖还原酶抑制活性等生物活性的研究进展进行了综述。     

乳酸菌发酵黄浆水的发酵特性以及其对大豆异黄酮的生物转化能力研究

目的 研究从中国传统发酵食品中筛选得到的9株植物乳杆菌发酵黄浆水的发酵特性以及其对大豆异黄酮的生物转化能力。方法 首先检测了黄浆水的基本成分,其次通过活细胞计数,pH和滴定酸度的检测探讨了乳酸菌发酵黄浆水的能力,之后采用吸光度法和高效液相色谱法分别检测了β-葡萄糖苷酶的酶活和6种大豆异黄酮的含量变化。结果 黄浆水中含有乳酸菌生长必需的营养物质，9株植物乳杆菌在黄浆水中都能够生长，同时能够产β-葡萄糖苷酶，具有一定的大豆异黄酮生物转化能力。其中，Lactiplantibacillus plantarum 17-17为最优良的菌株，该菌株在发酵6 h时活菌数可达8.49 lg CFU/mL，β-葡萄糖苷酶酶活可达0.68 U/mL，大豆苷元、大豆黄素和染料木素的含量分别增加了15.04、2.35和28.71 mg/L。结论 L. plantarum 17-17菌株可以有效提高黄浆水中大豆异黄酮的生物利用度，为黄浆水的高效回收利用提供了重要的理论依据。     

微生物酪氨酸解氨酶的研究进展

微生物转化生产次生代谢物质是重要的方式,但已知的微生物中未发现次生代谢物质的前体物质肉桂酸-4-羟基化酶(C4H)和香豆酸,这限制了微生物转化生产一些次生代谢产物。酪氨酸解氨酶(TAL)能跳过C4H将L-酪氨酸(L-Tyr)转化为香豆酸,为此对微生物来源的TAL的酶学性质、晶体结构、催化机制及TAL基因工程方面的研究进行了阐述,发现了TAL研究中存在的问题并初步提出了解决思路。     

微生物发酵法转化大豆异黄酮研究进展

大豆异黄酮是大豆中重要的生物活性物质,具有抗氧化、抑癌、类雌激素效应等多种活性功能,但在自然界中多以活性较低的结合态的糖苷存在,高活性的游离态的苷元含量却极低,需要进行转化以提高其活性。与酸水解、碱水解、Smith水解等一般转化方法相比,微生物发酵法将大豆异黄酮由糖苷型转化为苷元型,具有无污染、水解酶活性高、成本低等优点。该文对各种微生物转化大豆异黄酮的发酵效果及最佳发酵工艺和产酶条件进行综述和比较,以期为提高大豆异黄酮生物利用研究及其在食品、医药等行业中的应用提供参考。     

β-葡萄糖苷酶以及益生菌生物转化大豆异黄酮糖苷的研究进展

大豆及其加工制品中以糖苷形式存在的大豆异黄酮不利于人体的消化吸收,难以发挥其生物保健作用。本文综述了利用β-葡萄糖苷酶生物转化大豆异黄酮糖苷以提高生物利用率的国内外研究进展情况,重点介绍了外源性β-葡萄糖苷酶和产β-葡萄糖苷酶益生菌对大豆异黄酮糖苷的转化作用。     

大米蛋白酶解物—大豆苷元复合纳米粒子的构建

以大米蛋白为原料,经胰蛋白酶酶解后制备纳米载体,以大豆苷元为输送对象,构建了大米蛋白酶解物—大豆苷元的纳米输送体系,通过试验考察了大米蛋白水解度、大米蛋白酶解物质量浓度与大豆苷元质量浓度对制备的复合纳米粒子性质的影响。在大米蛋白水解度为12%,酶解物质量浓度为7.5 mg/mL,大豆苷元质量浓度为0.75 mg/mL的条件下制备的大米蛋白酶解物—大豆苷元复合纳米粒子,其平均粒径449.71 nm、多分散系数0.23,Zeta-电位-22.30 mV,稳定性较好;大豆苷元的包封率为60.07%,荷载量为6.01 mg/g,证明大米蛋白酶解物具有构建纳米载体荷载大豆苷元的可行性。     

酶解制备苷元型大豆异黄酮

以大豆异黄酮糖苷为原料,酶解制备苷元型大豆异黄酮。以水解率和苷元得率为指标对几种来源的β-葡萄糖苷酶、β-半乳糖苷酶、纤维素酶进行筛选,确定最适酶解用酶。通过单因素实验对酶添加量、底物质量浓度、酶解温度、pH、酶解时间进行优化。结果表明,最佳酶解工艺条件为：采用β-葡萄糖苷酶（300 U/g）,酶添加量7%,底物质量浓度1.6 mg/mL,酶解温度56 ℃,pH 4.8,酶解时间6 h。在最佳工艺条件下,大豆异黄酮糖苷的水解率及苷元得率分别达到96.84%和99.74%。     

小麦麸皮内源性β-木糖苷酶的酶学特性

以硝基苯酚-木糖苷为底物,在底物浓度为1 mg／ml ,提取液pH值为5的条件下,测定了小麦麸皮内源性β-木糖苷酶的酶活力,并对4种小麦麸皮内源性β-木糖苷酶的酶学特性进行了比较。结果表明：不同品种小麦麸皮内源性β-木糖苷酶的酶活力不同,同种小麦的粗麸皮和细麸皮中的内源性β-木糖苷酶的酶活力也不同;细麸皮中β-木糖苷酶的酶活力均高于粗麸皮中β-木糖苷酶酶活力,粗麸皮中郑麦8998的β-木糖苷酶酶活力最大,细麸皮中花培8号小麦β-木糖苷酶酶活力最大;郑麦8998、花培8号、郑麦366、郑麦90234种小麦粗麸皮内源性β-木糖苷酶的最适反应温度分别为60、50、50和60℃;最适反应pH值均为5;花培8号和郑麦366两种小麦粗麸皮内源性β-木糖苷酶在20～50℃范围内,酶活力稳定性较好,而郑麦8998和郑麦9023两种小麦粗麸皮内源性β-木糖苷酶在20～60℃范围内,温度对β-木糖苷酶的酶活力影响较小;4种小麦粗麸皮内源性β-木糖苷酶在pH值为5～6时,酶活力稳定性较好;Ca2＋、Co2＋促进β-木糖苷酶的酶活力,而Ag＋对β-木糖苷酶的酶活力有较强的抑制作用。     

大豆异黄酮糖苷酶的发酵和纯化

大豆异黄酮糖苷酶可水解大豆异黄酮的糖基使其生成苷元,从而提高大豆异黄酮的生物活性。实验中筛选得到了大豆异黄酮糖苷酶产酶菌株 Absidia sp．R,确定了该菌株发酵产酶培养基。采用硫酸铵沉淀和离子交换法对该酶进行了纯化,并初步确定了其分子质量。     

食醋浸泡对大豆中异黄酮转化的影响

采用红曲老醋浸泡大豆,对大豆自身的内源性β-葡萄糖苷酶(β-glucosidase,E.C.3.2.1.21,BG)促进大豆中异黄酮的转化进行研究。用合成底物p-硝基苯基-β-D-吡喃葡萄糖苷测定大豆BG酶活,高效液相色谱法测定大豆异黄酮的含量。结果表明,随着浸泡时间的延长,大豆BG酶活呈先上升后下降的趋势,浸泡4 h时酶活达到最高为0.74 U/g。在内源BG酶解和酸解的共同作用下,结合型异黄酮的糖苷逐渐被水解,其中丙二酰基葡萄糖苷型和β-葡萄糖苷型含量显著下降,分别降低47.39%和36.27%;苷元含量显著上升,增加12倍。食醋浸泡有利于提高大豆的生物效价。     

Metabolism of dietary soy isoflavones to equol by human intestinal microflora--implications for health

Soy isoflavones have received considerable attention. Individuals with isoflavones-rich diets have significantly lower occurrences of cardiovascular disease, osteoporosis, and some cancers. The clinical effectiveness of soy isoflavones may be a function of the ability to biotransform soy isoflavones to the more potent estrogenic metabolite, equol, which may enhance the actions of soy isoflavones, owing to its greater affinity for estrogen receptors, unique antiandrogenic properties, and superior antioxidant activity. However, not all individuals consuming daidzein produce equol. Only approximately one-third to one-half of the population is able to metabolize daidzein to equol. This high variability in equol production is presumably attributable to interindividual differences in the composition of the intestinal microflora, which may play an important role in the mechanisms of action of isoflavones. But, the specific bacterial species in the colon involved in the production of equol are yet to be discovered. Therefore, future researches are aimed at identifying the specific bacterial species and strains that are capable of converting daidzein to equol or increasing equol production.     

Food Microorganisms that Effectively Hydrolyze O-Glycoside but Not C-Glycoside Isoflavones in Puerariae Radix

ABSTRACT: Puerariae radix (PR) is known to contain abundant glucoside conjugates of daidzein isoflavones. This study was performed to determine the daidzin hydrolysis activity of the various strains of probiotic bacteria and edible fungi during fermentation of PR. Among the strains tested, Bifidobacterium sp. Int-57 showed the greatest β-glucosidase activity and daidzin hydrolysis activity. When yeast extract (2.5%) was added to the PR medium during fermentation with Bifidobacterium sp. Int-57, all of the daidzin was hydrolyzed to near completion. The addition of skim milk and whole milk also improved the conversion of daidzin into daidzein. Puerarin, a C-glucoside of daidzein in PR, was not hydrolyzed into daidzin by any of the experimental strains during fermentation. This study demonstrates that daidzin can be efficiently converted into daidzein by the proper combination of probiotic strains and fermentation conditions.     

Production of equol from daidzein by gram-positive rod-shaped bacterium isolated from rat intestine

Isoflavones (mainly daidzein and genistin) belong to the flavonoid group of compounds and are classified as phytoestrogens. In the intestine, daidzin is converted to daidzein by beta-glucosidase, and then daidzein is converted to O-desmethylangolensin (O-DMA) or equol via dihydrodaidzein by enzymes of intestinal bacteria. We isolated, for the first time, an anaerobic gram-positive rod-shaped strain capable of producing equol from daidzein. Its 16S rDNA gene sequence (1428 bp) showed 99% similarity with that of the human intestinal bacterium SNU-Julong 732 (AY310748) and 93% similarity with that of Eggerthella lenta ATCC 25559(T) (AF292375). This strain converted daidzein to equol via dihydrodaidzein in an equol-assay medium anaerobically. The addition of butyric acid and arginine increased the conversion ratio of daidzein to equol 4.7- and 4.5-fold, respectively.     

Effects of dietary daidzein supplementation of pregnant sows on carcass and meat quality and skeletal muscle cellularity of the progeny

The effects of dietary daidzein during late gestation on maternal performance, neonatal body composition, carcass and meat quality at market weight, and skeletal muscle cellularity were studied. Multiparous sows received a soybean-free diet (n=8, control) or the same diet supplemented with 1mg daidzein/kg body weight (n=7) daily from d 85 of gestation to parturition. Litter size, litter weight, and birth weight remained unaffected by daidzein feeding. In newborn piglets from litters >15, the proportions of muscle tissue and skin tended to be decreased (P=0.09) or increased (P=0.03), respectively, after gestational daidzein feeding. The body fat percentage was higher in response to maternal daidzein (P=0.04). Postnatal growth and carcass composition at 180d of age were not affected, however, maternal daidzein supplementation led to increases in longissimus muscle pH(45) (P=0.02) and pH(end) (P=0.11) in pigs from large litters. The proportion of fast-twitch glycolytic fibres in semitendinosus muscle was increased (P=0.06) by maternal daidzein feeding. The results suggest that supplemental daidzein in the maternal diet during late gestation marginally affects meat quality and skeletal muscle cellularity of the progeny.     

Bioavailability of soy isoflavones in rats Part I: application of accurate methodology for studying the effects of gender and source of isoflavones

There are limited and controversial reports about the effects of gender and source of isoflavones on their bioavailability. Moreover, several previous studies have not used appropriate methodology to determine the bioavailability of soy isoflavones, which requires comparing the area under the plasma concentration-time curve after both oral and intravenous injection (IV) administration. Therefore, the present study was conducted to determine the bioavailability of isoflavones from different sources following both oral and IV administration in male and female rats. Three sources of isoflavones; Novasoy (a commercial supplement), a mixture of synthetic aglycones (daidzein, genistein and glycitein) and a mixture of synthetic glucosides (daidzin, genistin and glycitin) were tested. Following administration, blood samples were collected at several time points (0, 10, 30 min and 1, 2, 8, 24, 48 h post oral gavage and 0, 10, 30, 45 min and 1, 2, 3, 4, 8 h post-IV dosing) and plasma isoflavones were measured by LC/MS. Bioavailability values for daidzein, genistein and glycitein were significantly (p <0.05) higher (up to sevenfold) in Novasoy and the glucoside forms of isoflavones compared with those of the aglycone forms. Moreover, significant (p <0.05) gender differences in the bioavailability of 7-hydroxyl-3-(4'-hydroxyphenyl)-chroman (a metabolite of daidzein), glycitein and daidzein were observed for Novasoy, with higher values in male rats. In summary, the source of isoflavones and the sex of rats had significant effects on isoflavone bioavailability.     

Escherichia coli biotransformation of daidzein fermentation products from soy‐based foods—relevance to food oestrogenicity‐based functionality

Oestrogenic isoflavones (e.g. daidzein) present in soy‐based foods are likely to be important in food functionality. Biotransformation of daidzein during fermentation‐based soy‐containing food manufacture forms three trihydroxy metabolites, 6‐ortho‐hydroxydaidzein (6‐OHD), 8‐ortho‐hydroxydaidzein (8‐OHD) and 3′‐ortho‐hydroxydaidzein (3′‐OHD) which alters the oestrogenicity of the final food product. We report that Escherichia coli (a key component of the gut microbiome) metabolism converts 8‐OHD to 6‐OHD with a likely concomitant increase in oestrogenicity. This means that the functionality of 8‐OHD‐containing soy‐based foods is altered by gut microbiome metabolism. This change in oestrogenicity‐based food functionality might have benefits for postmenopausal women, while being a feminising health risk for males and could add to the risk of oestrogen‐mediated precocious puberty in girls.     

Microbiological and chemical characterisation of ting, a sorghum-based sourdough product from Botswana

Ting is a fermented sorghum product of Botswana which is used to prepare porridges of different consistencies. This study aimed to characterise ting with respect to fermentation microbiota and metabolite composition, and to develop starter cultures for ting fermentation. The pH values of 10 ting samples from Botswana ranged between 3.5 and 4.0 and cell counts ranged between 1.2×10(8) and 1.2×10(10)cfu/g lactic acid bacteria. Yeast cell counts were below 10(5)cfu/g in all samples and Enterobacteriaceae were less than 10 cfu/g. The microbiota of samples consisted of 2-4 dominant strains and strains were identified based on the sequence of their 16S rRNA genes as Lactobacillus reuteri, Lactobacillus fermentum, Lactobacillus harbinensis, Lactobacillus plantarum, Lactobacillus parabuchneri, Lactobacillus casei and Lactobacillus coryniformis. Binary strain combinations were employed as starter cultures to produce ting. Major fermentation products were lactate, acetate and ethanol, additionally, 1,2 propanediol and 1,3 propanediol were produced by some strains. All strains were capable of producing ting, which was accompanied by a decrease in pH from 6.3 to 4 within 8h. Traditionally processed ting requires 2 to 3 days to attain a pH below 4; the starter cultures thus reduced the fermentation time to 8h.     

Combination of low dose of genistein and daidzein has synergistic preventive effects on isogenic human prostate cancer cells when compared with individual soy isoflavone

The reduced incidence of prostate cancer (PCa) in Asia countries has been attributed to high soy diets, and major soy isoflavones, in particular daidzein and genistein, are thought to be the source of the beneficial and anti-cancer effects of soy foods. However, attention has been drawn to the safety of using high levels of soy isoflavones in humans, which is especially the concern for consumers taking regular soy isoflavone dietary supplements. The main objective of this study is thus to identify a soy isoflavone combination with lower levels of daidzein and genistein to be a more efficacious and safer chemo-preventive agent for PCa. The anticancer effects of daidzein and genistein, and their combinations on early-stage androgen-dependent PCa cells (LNCaP) and bone metastatic LNCaP-derivative PCa cells (C4-2B) were compared. Cells were treated with varying concentrations of daidzein, genistein (25–200 μM) or their combinations (25 or 50 μM) and cell proliferation, apoptosis, cell cycles and cellular uptakes of the isoflavones were measured after 48 h. Daidzein and genistein showed a synergistic effect on inhibiting cell proliferation and inducing apoptosis of both PCa cells. Twenty-five μM daidzein/50 μM genistein and 50 μM daidzein/50 μM genistein significantly increased the apoptotic effects on C4-2B cells although they did not show any effect when used individually. Except 50 μM daidzein/50 μM genistein, all other combinations had no impacts on cell cycles. For treatment with soy isoflavone combination, genistein was always better taken up than daidzein by both LNCaP and C4-2B cells.     

Isoflavone phytoestrogen degradation in fermented soymilk with selected beta-glucosidase producing L. acidophilus strains during storage at different temperatures

Soymilk fermented with 3 selected Lactobacillus acidophilus strains were stored at various temperatures (-80 degrees C, 4 degrees C, 25 degrees C and 37 degrees C) for 8 weeks and the concentration of isoflavones determined weekly using RP-HPLC. The decreasing concentration of isoflavones in soymilk during storage due to degradation was found to fit the first order kinetics model. Isoflavone aglycones as well as isoflavone glucosides largely appeared to be stable during storage (P<0.01). Interestingly, the aglycone forms showed much smaller degradation as compared to glucoside forms at all the storage temperatures studied. Of the isoflavone aglycones, daidzein was found to be the most stable followed by genistein, while glycitein was least stable. Isoflavone aglycones such as glycitein, daidzein and genistein showed smaller degradation constants in fermented soymilk at lower storage temperatures (-80 degrees C and 4 degrees C) and higher degradation constants at higher storage temperatures (25 degrees C and 37 degrees C) with each strain. In contrast, glucosides glycitin and daidzin showed higher degradation at lower storage temperatures (-80 degrees C and 4 degrees C) and lower degradation at higher storage temperatures (25 degrees C and 37 degrees C). Storage temperature was therefore found to be very important in regulating the rate of degradation soy isoflavones in fermented soymilk.